Mammalia

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    Protocols in Current Issue
    Isolation and Culture of Single Myofiber and Immunostaining of Satellite Cells from Adult C57BL/6J Mice
    Authors:  Shujie Chen, Hongrong Ding, Xiangping Yao and Liwei Xie, date: 07/20/2019, view: 548, Q&A: 0
    [Abstract] Myofiber isolation followed with ex vivo culture could recapitulate and visualize satellite cells (SCs) activation, proliferation, and differentiation. This approach could be taken to understand the physiology of satellite cells and the molecular mechanism of regulatory factors, in terms of the involvement of intrinsic factors over SCs ...
    Cell Microencapsulation and Cryopreservation with Low Molecular Weight Hyaluronan and Dimethyl Sulfoxide
    Authors:  H. Gurruchaga, L. Saenz del Burgo, G. Orive, R. M. Hernandez, J. Ciriza and J. L. Pedraz, date: 02/20/2019, view: 1087, Q&A: 0
    [Abstract] Cryopreservation is commonly used for the storage of cells, tissues, organs or 3D cell-based products using ultra-low temperatures, which involves the immersion in liquid nitrogen for their long-term preservation. The cryopreservation of several microencapsulated cells is usually performed by the slow freezing with the dimethyl sulfoxide (DMSO) as ...
    Insulin Tolerance Test under Anaesthesia to Measure Tissue-specific Insulin-stimulated Glucose Disposal
    [Abstract] Insulin resistance is a pathophysiological state defined by impaired responses to insulin and is a risk factor for several metabolic diseases, most notably type 2 diabetes. Insulin resistance occurs in insulin target tissues including liver, adipose and skeletal muscle. Methods such as insulin tolerance tests and hyperinsulinaemic-euglycaemic ...
    Microarray, IPA and GSEA Analysis in Mice Models
    [Abstract] This protocol details a method to analyze two tissue samples at the transcriptomic level using microarray analysis, ingenuity pathway analysis (IPA) and gene set enrichment analysis (GSEA). Methods such as these provide insight into the mechanisms underlying biological differences across two samples and thus can be applied to interrogate a variety ...
    Transmission Electron Microscopy for Analysis of Mitochondria in Mouse Skeletal Muscle
    Authors:  Joseph D. McMillan and Michael A. Eisenback, date: 05/20/2018, view: 5369, Q&A: 0
    [Abstract] Skeletal muscle is the most abundant tissue in the human body and regulates a variety of functions including locomotion and whole-body metabolism. Skeletal muscle has a plethora of mitochondria, the organelles that are essential for aerobic generation of ATP which provides the chemical energy to fuel vital functions such as contraction. The number ...
    Electrophysiological Recordings of Evoked End-Plate Potential on Murine Neuro-muscular Synapse Preparations
    Authors:  Giulia Zanetti, Samuele Negro, Aram Megighian and Marco Pirazzini, date: 04/20/2018, view: 3056, Q&A: 0
    [Abstract] Neuromuscular junction (NMJ) is the specialized chemical synapse that mediates the transmission of the electrical impulse running along motor neuron axons to skeletal muscle fibers. NMJ is the best characterized chemical synapse and its study along many years of research has provided most of the general knowledge of synapse development, structure ...
    Mouse Satellite Cell Isolation and Transplantation
    [Abstract] Satellite cell (SC) transplantation represents a powerful strategy to investigate SC biology during muscle regeneration. We described here a protocol for SC isolation from green fluorescent protein (GFP)-expressing mice and their transplantation into murine muscles. This procedure was originally used to assess the effects of the hormone unacylated ...
    Biochemical Isolation of Myonuclei from Mouse Skeletal Muscle Tissue
    Authors:  Alicia A Cutler, Anita H Corbett and Grace K Pavlath, date: 12/20/2017, view: 4849, Q&A: 0
    [Abstract] Skeletal muscle provides the contractile force necessary for movement, swallowing, and breathing and, consequently, is necessary for survival. Skeletal muscle cells are unique in that they are extremely large cells containing thousands of nuclei. These nuclei must all work in concert to maintain skeletal muscle function and thereby maintain life. ...
    Muscle Histology Characterization Using H&E Staining and Muscle Fiber Type Classification Using Immunofluorescence Staining
    Authors:  Chao Wang, Feng Yue and Shihuan Kuang, date: 05/20/2017, view: 10781, Q&A: 0
    [Abstract] Muscle function is determined by its structure and fiber type composition. Here we describe a protocol to examine muscle histology and myofiber types using hematoxylin and eosin (H&E) and immunofluorescence staining, respectively. H&E stain nucleus in blue and cytoplasm in red, therefore allowing for morphological analyses, such as myofiber ...
    Isolation, Culturing, and Differentiation of Primary Myoblasts from Skeletal Muscle of Adult Mice
    Authors:  Lubna Hindi, Joseph D. McMillan, Dil Afroze, Sajedah M. Hindi and Ashok Kumar, date: 05/05/2017, view: 13065, Q&A: 4
    [Abstract] Myogenesis is a multi-step process that leads to the formation of skeletal muscle during embryonic development and repair of injured myofibers. In this process, myoblasts are the main effector cell type which fuse with each other or to injured myofibers leading to the formation of new myofibers or regeneration of skeletal muscle in adults. Many ...



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